Description: The overall goal of this revised SBIR application is to produce and test monoclonal antibodies (mab) to human CD40 that possess "super-agonistic" activity, as part of an adjuvant platform. The term "superagonist" is defined by specific immunological activities that these mabs possess, activities that are distinct from and superior to those of current CD40 agonists. At this time, anti-murine CD40 superagonists exist, however their anti-human (h) CD40 counterparts do not exist. ImmuRx is committed to the development of "super-agonistic" anti-hCD40 mabs as part of a novel, proprietary vaccine adjuvant platform. This adjuvant will be an integral component of therapeutic anti-tumor immunity. This proprietary adjuvant platform is based on the combined use of CD40 "superagonists" and TLR agonists to amplify cell-mediated immunity effectively and safely. The goal here is to produce and identify lead CD40 "superagonists" for continued development into a Phase II program. "Super-agonistic" mabs specific for hCD40 will be identified and produced through a modified Phage display approach. Under contract with Alligator BioSciences, a human H-L molecular library will be produced from 8 previously identified agonistic anti-human CD40 mabs. From these 8 mabs, the VH/VL library will be mutated using the FIND proprietary technology allowing millions of new variants to be produced and thousands to be tested. An additional round of affinity/functional maturation will be performed. Alligator will provide 40-100 mabs to hCD40 with high levels of agonistic activities. These will represent the starting library of mabs for the SBIR from which lead super-agonistic mab candidates will be tested and selected. In addition, ImmuRx will also take a conventional, independent approach and produce mouse anti-h CD40 mabs. While "super- agonists" will be identified by their in vitro activities, definitive verification of the activities of lead candidates and their ability to synergize with TLR agonistics will be determined by in vivo testing in hCD40 knock-in mice. These "humanized" knock-in mice will allow the testing of lead CD40 superagonists for inducing heightened cell-mediated immunity and the assessment of possible toxicity due to antibody-based therapy, prior to advancing into sub-human primates. There is a substantial need for potent and safe adjuvants to immunize against melanoma. Super-agonistic anti-CD40 mabs represent part of a powerful and new adjuvant platform that will allow the development of such vaccines. PUBLIC HEALTH RELEVANCE: Vaccines have been the most successful interventional therapy that mankind has known. They have all but wiped certain diseases from the face of the earth. This application is to produce a drug that will enhance the power of vaccines to improve their capacities to treat metastatic melanoma.